纹枯菌GXE4菌株所携带的真菌质粒pGXRS-1和-2序列克隆及分析.docVIP

  纹枯菌 GXE4 菌株所携带的真菌质粒 pGXRS-1 和-2 序列克隆及分析# * 5 10 15 20 25 30 35 40  摘要：前期研究自广西桂林稻田中分离获得了一株致病力衰退的菌株 GEX4，证实其携带有 独立于染色体外遗传因子 3.6-kb frags。本论文通过对该因子的物理特性及其序列克隆分 析，获得以下结论。DNase I、RNase、限制性内切酶、外切酶 III 等降解试验表明 3.6-kb frags 具双链 DNA 特性，且 3’端不含有发夹结构或蛋白质保护性结构。对 3.6-kb frags 进行了 克隆和序列分析，发现在 3.6-kb frags 至少包含有两个大小类似、序列具有同源性的真菌 质粒，分别命名为 pGXRS-1 和 pGXRS-2；pGXRS-1 没有氨基酸数超过 110 的阅读框，推定不 具有编码功能；不完全序列分析表明 pGXRS-2 也没有较大的阅读框。DNA 杂交结果显示位于 约 23 kb 和 7.0 kb 处还存在与 pGXRS 具明显杂交信号的 DNA 片段。根据这些研究结果初步 推定 pGXRS-1 是一种新线性真菌质粒。 关键词：纹枯菌；真菌质粒；质粒属性；克隆分析 中图分类号：S436.63 Cloning and analysis of fungal plasmids pGXRS-1 and -2 isolated from strain GXE4 of Rhizoctonia solani Xie Jiatao, Fu Yanping, Cheng Jiasen, Jiang Daohong (College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070) Abstract: It was previously reported that extra linear chromosmal element 3.6-kb frags exsited in Rhizoctonia solani strain GXE4, which was a hypovirulent fungal strain. In the present paper, the physical properties and sequence of this element were analysized and the results was as following. 3.6-kb frags was a kind of dsDNA and confirmed via digestion with DNase I、RNase and restriction enzyme, and no protein or knob-shaped structure in 3 end of 3.6-kb frags becaused 3.6-kb frags could be digested by exonuclease III. Sequence analysis of 3.6-kb frags revealed that there existed two size similar funal plasmids (named as pGXRS-1and pGXRS-2) at least and confirmed by DNA southern blot. The DNA of pGXRS-1 was presumed not to encode any protein longer than 110 amino acids. The DNA of pGXRS-2 did not have a longer ORF either. The results of genomic DNA southern blot showed that there was a significant hybridization signal with DNA fragments of 3.6-kb frags at 23 kb and 7.0 kb. Based on the results, pGXRS (-1 and -2) existed in the R. solani strain GXE4 may be two nevol type fungal plasmids. Key words: Rhizoctonia solani; fungal plasmid; physical property; sequnece analysis 0 引言 丝状真菌质粒一般呈线性，少数是环状，双